Topical compositions containing LYCD and other topically active medicinal ingredients for the treatment of ACNE

ABSTRACT

A topical composition comprising LYCD together with known topically active useful medicinal agents such as anti-wrinkling, antibiotic, anticancer, antifungal, antiinflammatory such as anti-acne, antiviral, wound healing, and hair-growing agents. The LYCD works together with the other active agents to achieve a synergistic result more effective than can be obtained from the topical agents individually, and more effective than could be predicted from the mere addition of the known efficacies of the individual ingredients.

RELATED APPLICATION

The present application is a continuation-in-part of application U.S.Ser. No. 07/394,862 filed 08/16/89, U.S. Pat. No. 4,942,031; which inturn is a continuation-in-part of application U.S. Ser. No. 07/159,390,filed Feb. 23, 1988, the latter application now abandoned, bothapplications being of the present inventor, Robert H. Levin.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to topically applied medicinalcompositions, and more particularly refers to such compositions havingactive topical medicinal ingredients, and additionally having LYCD inamounts sufficient to act with the other active ingredients to providesynergistic therapeutic results.

2. Description of the Prior Art

LYCD as utilized herein in the specification and claims is the acronymfor Live Yeast Cell Derivative. The material is also known as SkinRespiratory Factor (SRF), Tissue Respiratory Factor (TRF), andProcytoxoid (PCO). The product, LYCD, is an alcoholic extract of viableSaccharomyces cerevisiae. The material is produced and marketed by MDHLaboratories, Inc., Cincinnati, Ohio 45210 as a standard article ofcommerce. Another producer of LYCD is Universal Foods Corporation,Fermentation Division, Milwaukee, Wis. 53202. LYCD is available forexperimental use as a bulk drug assaying 5 units to 40 units/mg ofrespiratory activity. In topical medicinal preparations it ischaracterized and quantified in terms of Skin Respiratory Factor (SRF)units. A unit of activity is calculated as the amount of SRF which isrequired to increase the oxygen uptake of 1 mg of dry weight ratabdominal skin by 1 percent at the end of a 1 hour testing period in aWarburg apparatus.

LYCD is also available as LYCODERM® ointment containing 2,000 units SkinRespiratory Factor (SRF) per ounce, from Arel Pharmaceuticals, Inc.,Cincinnati, Ohio. In the prior art the well know hemorrhoidal ointment,PREPARATION H®, contains 2000 units of SRF (ca 1%) per ounce ofointment.

J. Z. Kaplan (Arch. Surge. 119(9) p. 1005-8 (1984) has reported that, ina double blind human skin graft study donor sites treated with LYCDointment, statistically significant earlier angiogenesis andepitheliazation occurred as compared with donor sites in the samepatients treated with ointment bases (without LYCD). This studyconfirmed earlier laboratory reports such as that of Wm. Goodson et. al.Journal of Surgical Research 21: 125-129 (1976) showing that LYCD iscapable of stimulating wound oxygen consumption, epitheliazation, andcollagen synthesis.

As reported in the Cincinnati Inquirer of Dec. 12, 1986, Ashlley HunterCosmetic Co. offers a facial cream containing LYCD to minimize wrinkles.

For milder forms of acne, which may be inflammatory, topical benzoylperoxide (BP), an antibacterial and oxidizing agent, topicalerythromycin (EM), clindamycin phosphate (CP), oral tetracyclines, or EMantibiotics are usually effective treatments, as disclosed in the priorart. (C. D. Bunker, Drugs Today, 24, 229 (1988).

SUMMARY OF THE INVENTION

It is an object of the present invention to provide topically appliedpharmaceutical compositions suitable for the treatment of variousailments and physical conditions of the skin such as acne, bed sores,burns, infections, trauma, ulcers, wounds, and wrinkles.

It is a further object to provide compositions of the type describedwhich are more effective than compositions presently known in the art.

It is a prime object of the invention to provide topical compositions ofthe type described for the treatment of acne, and more particularly,severe forms of acne, which compositions are more effective as remediesthan the compositions presently known and used in the art.

The foregoing and other objects, advantages and characterizing featureswill become apparent from the following description of certainillustrative embodiments of the invention.

According to the invention, pharmaceutical compositions for topicalapplication are provided by utilizing LYCD in combination with knownpharmaceutical agents or remedies. The LYCD acts synergistically withthe other agents to provide a composition having greater effectivenessthan that of the individual agents, and a greater effectiveness thancould be predicted by combining (in an additive fashion) the known ortheoretical effectiveness of the individual ingredients.

According to the invention, it has been further found that many patientswith severe acne, refractory to even long term treatment with a varietyof the conventional acne remedies experience significant improvement intheir acne condition within thirty days when treated twice daily with acombination of any of the conventional antibacterial acne medicationsand LYCD.

It has been additionally found that mild to moderately severe acne canbe treated and the condition ameliorated by the application of a topicalpharmaceutical composition comprising LYCD in a suitable vehicle, evenin the absence of conventional acne remedies.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

LYCD, Live Yeast Cell Derivative, also known as SRF or TRF, is acommercial material produced by the method set forth in U.S. Pat. Nos.2,239,345, 2,320,478, and 2,320,479. which are herein incorporated byreference, and is standardized as units with 1 unit (U) of SRFincreasing the uptake of oxygen by minced rat abdominal skin (1 mg. dryweight) by 1% in a 1-hour measurement by Warburg manometry.

Example 1

The compositions of the invention may be produced by either of twogeneral methods. In the first method, for example, an ointmentcomposition may be formulated by mixing LYCD with conventionalointment-forming ingredients. One such ointment composition isLYCODERM®. a registered trademark material marketed by ArelPharmaceuticals, Cincinnati, Ohio, having the following compositions:

    ______________________________________                                                          Per 100 Parts                                               ______________________________________                                        Beeswax             4.0                                                       Lanolin             4.0                                                       Petrolatum          87.9                                                      Shark liver oil     3.0                                                       Phenyl mercuric nitrate                                                                           0.01                                                      LYCD (2000) SRF units/ounce                                                                       1.0 approx.                                               Thyme oil           0.1                                                       ______________________________________                                    

Procedure

In preparing the above compositions, the beeswax, petrolatum, sharkliver oil and phenyl mercuric nitrate are treated together to atemperature of 140 deg. F. in a steam-jacketed s.s. kettle. Thematerials are mixed until the mixture is uniform. Then the steam isturned off and cooling water is introduced while mixing is continued.When the mixture has reached a temperature of 110 deg. F., the LYCD andthyme oil are added. When the composition becomes uniform, mixing isstopped. Filling may be carried out at temperatures of between 90 and110 deg. F.

In utilizing the first method of producing the compositions of theinvention, the LYCODERM® ointment as produced above but utilizing 200 to1800 SRF units per ounce of formulation is mixed with the knownpharmaceutical agent by a conventional method.

The second method of carrying out the present invention comprises mixingbulk LYCD into the conventional pharmaceutical composition.

In general, it has been found that effective compositions are formulatedusing LYCD concentrations in the range of about 0.1% to 0.5% by weightof the composition, constituting, for example, 10% to 50% of thatemployed in the basic LYCODERM® ointment formulation. This translatesinto 200 to 1000 LYCD or SRF units per ounce of combined product. Statedin another way, the new compositions have a LYCD concentration ofapproximately 0.1% to 0.5% compared with approximately 1.0% LYCD in thebasic LYCODERM® formulation. However, for certain compositions LYCD inthe amount of 0.8% of the total composition produces optimum results.

Correspondingly, the other active components of the new compositions arefound to be synergistically efficacious at concentrations in the rangeof 5% to 50% of that found to be efficacious in marketed pharmaceuticalproducts.

ANTI ACNE COMPOSITIONS Example 2

The above-described LYCODERM® ointment was formulated using 1000 unitsof S RF per ounce. To this formulation was added sufficient 2%erythromycin to yield an ointment containing 0.5% LYCD and 1%erythromycin. In a controlled clinical trial this composition wasequally effective in the topical control of acne vulgaris as aconventional 2% ointment preparation of erythromycin. (An example ofsuch a medicinal preparation, NDA 50-584, Akneymycin Topical Ointment,2%, is a topical erythromycin accepted by the FDA for topical control ofacne vulgaris.)

Example 3

Alternatively, a marketed anti-acne 2% Erythromycin product wasreformulated as follows:

    ______________________________________                                        Erythromycin Base    1%       w/v                                             LYCD                 0.5%     w/v                                             Alcohol              55%      v/v                                             Oleyl Alcohol        5%       w/v                                             Perfume              q.s.                                                     Propylene Glycol     to 100%                                                  ______________________________________                                    

Following the same treatment regimen, the 1% Erythromycin solutioncontaining approximately 0.5% LYCD of Example 2 (assaying 1,000 units ofSRF per ounce of formulation) was found to be more effective than asimilar 2% Erythromycin solution without LYCD in the treatment of Acnegrades 2 and 3 (moderate to severe).

Example 4

In an analogous fashion, Lincomycin Hydrochloride or ClindamycinPhosphate at a concentration of 0.5% is combined with 0.5% LYCD in theLYCODERM® formulation to provide an ointment as effective in treatingcommon acne as the conventional 1% Clindamycin Phosphate topicalmedication, and more effective than either agent individually orpredictable in their combination.

Example 5

Ten patients suffering with severe acne completely refractory to monthsof prior treatment with a variety of conventional acne medications weretreated with an admixtured combination of Cleocin T (clindamycinphosphate solution) and LYCODERM® Ointment. The final formulationcontained approximately 0.34% clindamycin and 0.56% LYCD. Each subjectapplied this formulation twice daily for 30 days with the followingresults:

Patient 1: female, age 16: Refractory to Acromycin, Erythrocin, diet,local treatment. The patient was subsequently treated with the low dosecombination product of Example 5, and as a result experienced 80%improvement.

Patient 2: male, age 25: Refractory to Achromycin, curattage, Acutane.The patient was then treated with the low dose combination product ofExample 5, and as a result experienced 90% improvement.

Patient 3: male, age 30: Refractory to proprietary medicines,Erythromycin, Achromycin, Fostex (10% Benzoyl Peroxide). The patient wasthen treated with the low dose combination product of Example 5, andexperienced 50% improvement.

Patient 4: female, age 21: Refractory to astringents, Erythromycin, anddiet. The patient was then treated with the low dose combination productof Example 5, and experienced 85% improvement.

Patient 5: Female, age 18: Refractory to Achromycin, diet, andastringents. The patient was subsequently treated with the low doseformulation of Example 5, and experienced almost 100% improvement.

Patient 6: female, age 24: Refractory to Achromycin, curettage, anddiet. The patient was subsequently treated with the low dose formulationof Example 5, and experienced 85% improvement.

Patient 7: Female, age 24: Refractory to Achromycin, diet, local Rx, andproprietary drugs. The patient was subsequently treated with the lowdose formulation product of Example 5, and experienced 85% improvement.

Patient 8: Female, age 29: Refractory to Vitamin A ointment, diet, andAchromycin. The patient was subsequently treated with the low doseformulation of Example 5, and experienced 80% improvement.

Patient 9: Male, age 30: Refractory to Achromycin, local Rx, andAcutane: The patient was subsequently treated with the low doseformulation of Example 5, and experienced 80% improvement.

Patient 10: Female, age 20: Refractory to Achromycin, and coal tarproducts. The patient was subsequently treated with the low doseformulation of Example 5, and experienced 85% improvement.

Example 6

In an analogous fashion topical acne treatment formulations containing5% or 10% Benzoyl Peroxide anti-acne formulations were admixed withLYCODERM® ointment to provide lower dose Benzoyl Peroxide/LYCDcombination products more effective than the higher concentration ofBenzoyl Peroxide conventionally used in the treatment of acne vulgaris.Additionally, desquamation, symptoms of burning, and other side effectswere less frequent.

Example 7

In a further test 1.5 oz. FOSTEX® 5% Benzoyl Peroxide Gel was admixedwith 2 oz. LYCD ointment to provide a formulation containingapproximately 2% Benzoyl Peroxide and 0.5% LYCD (assaying approximately1,000 units of SRF per ounce of formulation). Further 1 oz. ofCLEARASIL®, 10% benzoyl peroxide anti-acne cream, was admixed with 4 oz.of LYCODERM® ointment to provide a formulation containing approximately2% benzoyl peroxide and 0.8% LYCD. Both compositions are effective intreating mild to severe acne.

Example 8

A LYCODERM® composition is prepared comprising containing 1000 units ofSRF per ounce (ca 0.5%) and 0.25% of vitamin A acid to provide a topicalointment composition more effective than the retinoic acid by itself inameliorating the effects of moderate to severe acne.

Example 9.

The LYCODERM® formulation was applied for twelve weeks to mild tomoderately severe acne afflicted subjects in the absence of conventionaldrugs used in the treatment of acne. The treatment resulted in a 40%reduction in inflammatory lesions. For clinical testing purposes thesame formulation was prepared as a placebo by leaving out the LYCD.During this period the placebo, when applied to similarly afflictedpatients proved to be ineffective,

Example 10

Various protein/peptide growth factors have been used as topicalwound-repair agents. It has now been found that Epidermal Growth Factor(EGF), when used in concentrations of 0.0001% in a suitable topicalpharmaceutical formulation is an effective treatment for moderate acne.

Further, according to the method of the present invention, compositionscontaining 500 units per ounce of LYCD/SRF incorporated into commercialformulations of recombinant/human Epidermal Growth Factor preparationsor their reduction products (containing 0.0001% rh EGF) aresynergistically more effective in treating severe acne.

FIRST AID, BURN, AND WOUND-HEALING COMPOSITIONS Example 11

Several commonly used topical antibiotic preparations are accepted bythe FDA for non-prescription (OTC) use to help prevent infection and aidin the healing of minor cuts, burns, and abrasions. One such preparationcontains 0.5% neomycin sulfate. The second contains 500 units ofbacitracin, 5000 units of polymixin B sulfate, and 0.5% neomycin sulfateper gram of composition. To each of these formulations was added 1000units of LYCD (approximately 0.5%) per ounce of product. The resultingLYCD antibiotic compositions were synergistically more effective fortopical first aid use than the conventional components when usedseparately.

Example 12

In an analogous example, a LYCODERM®/antibiotic combination ointmentproduct was formulated as follows:

    ______________________________________                                        Polysorbate 80  2        pounds   1.00% w/w                                   Polymixin B (7,700 iu/mg)                                                                     117.02   grams    0.129%                                                                              w/w                                   Bacitracin Zinc (690 u/mg)                                                                    656.81   grams    0.724 w/w                                   Phenyl Mercuric Nitrate                                                                       9        grams    0.01% w/w                                   LYCD (12 u/mg)  266.7    grams    0.50% w/w                                   Deionized water 4# 7     oz       2.50% w/w                                   Beeswax (white) 8# 1     oz       4.00% w/w                                   Lanolin         8# 1     oz       4.00% w/w                                   Petrolatum      167# 13  oz       83.93%                                                                              w/w                                   Shark liver oil 6# 1     oz.      3.03% w/w                                   Thyme oil       90       grams    0.10% w/w                                   ______________________________________                                    

Procedure

In preparing the above composition, the first four ingredients weremixed and suspended well. The LYCD is dissolved in the water and mixedwell with the first ingredients. The resulting solution was heated to140° F.

Separately, the third group of ingredients was mixed in a cleancontainer and heated to 160° F. With good mixing the first solution wasadded to the contents of the container; the thyme oil was then added andthe composition permitted to cool to 60°-80° for filling intocontainers.

The product thus produced is ideal for treating minor cuts, burns, andscratches. It appears to work rapidly and especially well on infants andchildren, where the ointment formulation functions somewhat like anocclusive dressing.

Example 13

A similar LYCODERM®/antibiotic ointment formulation was also preparedusing LYCD at a concentration of approximately 1.0% (2000 unitsSRF/ounce of product.

Used twice a day for two to four weeks as an emollient in a group of 10elderly patients suffering from bed sores, this product resulted in 50to 100% healing.

Separately, LYCD or LYCODERM® ointment may be incorporated into firstaid type bandages or dressings to provide a superior product for burnand wound healing.

Example 14

Topical antiinfective compositions for wounds including burns may beprovided utilizing LYCD in combination with silver sulfadiazine and asuitable ointment-forming base, or LYCD in combination withpovidone-iodine and a suitable ointment-forming base.

Live yeast cell derivative (LYCD) per se increases collagen formation.It is accepted in the art that most agents promoting experimental woundhealing appear to act primarily to promote collagen synthesis.Controlled clinical studies of LYCD have demonstrated both clinicallyand statistically earlier angiogenisis, initiation and completion ofepithelialazation, and acceleration of wound healing.

The use of a composition such as LYCODERM® formulated with 2000 units ofLYCD per ounce of ointment (about 1.0%), and also containing 3% of sharkliver oil has also been shown clinically to promote wound healing,including a range of first, second and third degree burn wounds.

Separately, a number of lymphokine/cytokine proteins have been foundwhich enhance wound healing by directly activating macrophages orindirectly stimulating the skin immune system. More than several dozenof these naturally occurring growth factors have been reported in theliterature, but are difficult to isolate and characterize, and mayactually overlap in identity. Therapeutic doses, although measured infractions of milligrams, are very costly.

Example 15

Epidermal Growth Factor (EGF) is one such growth factor which has beenused topically at a concentration of 0.0001% to accelerate normal woundhealing by 15-20 percent. According to the method of the presentinvention, compositions containing 500 units of LYCD as SRF(approximately 0.25%) and 0.0001% EGF are synergistically more effectivein treating chronic epidermal ulcers. Similarly using LYCD at 0.1% to0.5% concentrations (equivalent to 200-1000 LYCD units per ounce ofproduct) in compositions with Fibroblast Growth Factor (FGF),Platelet-Derived Growth Factor (PDGF), Transforming Growth Factor-alpha(TGF-alpha), Transforming Growth Factor-beta (TGF-beta), or Insulin-likeGrowth Factor-1 (IGF-1) has provided synergistic wound healingcompositions. Both partial and full incisional wounds show synergistichealing patterns. Compositions consisting of several of these growthfactors formulated with LYCD as in LYCODERM® also result insynergistically acting wound healing products.

Example 16

In the area of surgical incisions and wounds, as for example inGastrointestinal Surgery, excess scarring of the tissue is a notuncommon side effect resulting in surgical adhesions which may require asecond operation for correction.

It has now been found that a 10% sterile isotonic saline solution ofLYCD can be used as a final incision lavage in such surgical proceduresto minimize scarring during the wound healing process. A range of LYCDconcentrations of 1% to 25% may be used; and isotonic lactose as well assterile water are suitable vehicles for this purpose. Additionally, aspreviously disclosed herein, the LYCD solution stabilizes other growthfactor/lymphokine/cytokine proteins at the 1 to 100 nanogram per ml.concentrations required for therapeutic purposes. Thus, for example, 5nanograms per ml. each of insulin-like growth factor-1 and plateletderived growth factor are added to the 10% LYCD solution to make agrowth factor "cocktail" which, when used as a final lavage,significantly reduces the "adhesion" side effect of many cardiac,neurovascular, and gastrointestinal surgical procedures.

Separately as an adjunct to orthopedic surgery, LYCD, formulated as a 5%sterile solution is combined with one or more of a group of growthfactors/cytokines (in concentrations of one to 100 nanograms per ml.) toprovide compositions for the acceleration of healing of bone and otherhard tissue injury. Representative factors include: Bone MorphogenicProtein, Cartilage-derived Growth Factor, Cartilage Inducement Factor,Connective Tissue Activating Peptide III, Fibroblast GrowthFactor-basic, Osteogenic Growth Factor, Osteogenic Protein, Osteogenin,Skeletal Growth Factor, Tissue Inhibitor of Metalloproteinese,Transforming Growth Factor alpha, and Transforming Growth Factor beta.

Example 17

In another embodiment of this invention lymphokine/cytokine modulatingchemicals, such as Tilorone and its congeners, are formulated intotopical wound healing compositions in combination with LYCD. Thus theLYCODERM® formulation previously described is prepared using 1000 unitsof SRF per ounce (ca 0.5% and 0.1% of Tilorone to produce asynergistically effective ointment for the treatment of severe burnwounds and non-healing epidermal ulcers. Depending on the formulation,Tilorone synergy can be demonstrated at concentrations ranging from0.01% to 0.5% in epithelial tissue repair experiments.

ANTIFUNGAL COMPOSITIONS Example 18

Topical antifungal compositions are used in the treatment of cutaneousor mucocutaneous mycotic infections caused by Candida species,pathogenic dermatophytes, other yeasts, and various superficial fungalinfections of the skin. Tolnaftate USP at the level of 0.5%, whenformulated with LYCODERM® containing LYCD at a concentration of 1000units per ounce (ca 0.5%) provides a synergistic composition moreeffective than the standard topical antifungal preparation containing1.0% of Tolnaftate. In an analogous manner, Nystatin USP at aconcentration of 50,000 units per gram when formulated with LYCD at the0.5% concentration, and chlortrimazole 0.25% provided compositions equalin antifungal effectiveness with the topical antifungals used alone at 3and 4 times the concentration employed in the compositions of theinvention.

Example 19

In similar manner 30 cc. of clotrimazole solution, USP 1%, was admixedwith 3 oz. of LYCODERM® ointment, and the resulting combined formulationused to treat a severe tinea pedis infection in a 70 year old male. Withtwice a day application into the affected and surrounding skin areas,the infection cleared up within a week.

ANTIVIRAL COMPOSITIONS Example 20

Topical antiviral medicinal agents are licensed by the FDA particularlyfor the treatment of herpes simplex and herpes genitalis. The agentidoxuridine is marketed as a 0.5% ointment. When formulated as a 0.1%concentration of idoxuridine and 0.5% concentration of LYCD (containing1000 units SRF per ounce), the topical composition showed synergisticanti-herpes simplex viral activity. Similarly, a topical compositioncontaining 0.5% acyclovir and 0.5% LYCD (containing 1000 units of SRFper ounce of LYCODERM® ointment formulation) gave better results in thetreatment of herpes genitalia than would be anticipated from the knownantiherpes activities of acyclovir in the absence of LYCD. The antiviralefficacy of topical formulations of alpha, beta, or gamma interferons(used at doses of 3 million to 10 million IU) per 30 ml. solution arealso enhanced synergistically by the addition of 0.1% to 0.5% of LYCDrepresenting 200 to 1000 units of SRF per ounce of topical composition.

In laboratory cell culture studies, the effect of LYCD, lymphokines andlymphokine modulating chemicals on vaccinia virus infected monkey kidneyBSC-40 cells, or human epidermoid A431 cells was investigated. Vacciniavirus is a DNA virus, as is herpes.

Example 21

Pre-treated experiments: Each cell line was pretreated with a candidateantiviral compound for 24 hours, washed, and then infected with lowmultiplicities of vaccinia. After 24 hours of infection, virus wastitered on BSC-40 monolayers. The data is expressed in placque-formingunits (PFU); and the results of duplicate experiments averaged.

Post-infection experiments: Vaccinia infected cells were exposed to thetest material for 24 hours.

The 24 hour preincubation of BSC-40 cells with 100 to 200 micrograms perml. of LYCD, followed by vaccinia virus infection resulted in a 30% to40% reduction in PFU. Higher concentrations of LYCD did notquantitatively change the viral inhibition.

Example 22

The immunomodulating drug Tilorone, and two of its analogs, RMI-11567,and RMI-11645 were similarly tested for their ability to induce anantiviral state in BSC-40 cells by pretreatment of the cells for 24hours. (See Progress in Medicinal Chemistry, vol. 18, pp. 136-190, 1981for a description of these compounds) In these experiments 2-3micrograms per ml. of the tilorones resulted in an inhibition of viralgrowth which peaked at 40%, with RMI 11567 being the most active.

Example 23

In analogous experiments, pretreatment with the lymphokines gammainterferon, and Interleukin-I (IL-1) at even lower doses (250 units perml. and 10 units per ml., respectively) resulted in a 60% to 90%stimulation of viral growth. However, combination of each of thesematerials with 100 micrograms per ml. of LYCD synergistically reduced orreversed this stimulation.

Example 24

Surprisingly, in Post-infection cell culture experiments, 1 microgramper ml. of RMI-11567 and 100 micrograms per ml. of LYCD each separatelycaused a 30% stimulation in viral growth; however, the combination ofRMI-11567 and LYCD resulted, synergistically, in a 15% inhibition ofvaccinia growth as measured by plaque forming units (PFU). The percentstimulation/inhibition was calculated by comparing the PFU to concurrentcontrol experiments which did not contain any drug.

Example 25

In analogous Post-Infection studies with A431 cells, somewhat similarresults were obtained. Thus 8 (standard) units per ml. of alpha or betaInterleukin-I resulted in a slight (3%) inhibition of vaccinia growth asmeasured by PFU; and 100 micrograms per ml. of LYCD resulted in an 18%inhibition of PFU. The combination of 8 units per ml. of IL-1 and 100micrograms per ml. of LYCD resulted in a synergistic 32-35% inhibitionof viral growth as measured by placque-forming units (PFU).

The experiments described in the examples above demonstrate that in cellculture LYCD has significant antiviral activity which cansynergistically enhance the antiviral effects of some lymphokines andlymphokine modulating chemicals.

As more fully described below, a standardized LYCD preparation assaying12 units of Skin Respiratory Factor (SRF) per mg. was used in the abovedescribed cell culture studies.

Example 26

The addition of 0.5% fluorouracil to the above-described LYCODERM®ointment formulated with 1000 units of SRF (about 0.5% LYCD) per ounceof product provides a composition for the topical treatment of multipleactinic (solar) keratoses which is synergistically more effective than aconventional topical preparation formulated with 1.0% fluorouracil.

ANTIINFLAMMATORY COMPOSITIONS Example 27

Triethanolamine salicylate (10%) is formulated in lotions and creams toprovide a topical external analgesic agent for temporary relief fromminor pains of arthritis, rheumatism, and muscular aches. Whenformulated using 5% of triethanolamine salicylate and 200 to 1800 unitsSRF per ounce of product (equivalent to approximately 0.1% to 0.8% ofLYCD), the new composition was synergistically more effective than theoriginal analgesic product containing 10% of triethanolamine salicylate.In an analogous manner, when LYCODERM® is formulated with non-steroidalantiinflammatory agents such as ibuprofen or its isomers (at a level of1.0% to 5.0%), the resulting topical compositions have a higher level oftopical anti-inflammatory efficacy than that demonstrated by the samepreparation of the non-steroidal antiinflammatory agent (NAIA)formulated without LYCD.

The adreno-corticoid steroids demonstrate pleitropic activity in cellculture systems and as topical anti-inflammatory agents.

Metabolically, LYCD biological activity at the cellular level in animaland human skin results in an increase in oxygen respiration and cellgrowth. However, in specific human cell lines including humanfibroblasts, the effects are variable.

Example 28

A number of standard human cell lines were evaluated and methodology wasdeveloped leading to the use of A431 cells, Am Type Culture CollectionCLL 1555, as a suitable human cell line to study the effect of LYCD onoxygen respiration and cell growth. In order to facilitate study of themetabolic interaction of LYCD with lymphokines, cytokines, other growthfactors, and topically useful therapeutic agents, it was necessary toachieve reproducible base line results in a defined serum-free cellculture medium. Cell respiration was quantified using a Clark oxygenelectrode oxygraph apparatus. Experiments were done in duplicate, andcell number measurements were always done in quadruplicate, for anysingle experiment, the amount of (oxygen) respiratory stimulation orinhibition of a measured number of A431 cells can be correlated with theconcentration of added LYCD. Respiration is measured within 6-10 minutesof adding the LYCD and/or other substrates.

A standardized LYCD preparation assaying 12 units of SRF/mg. was used inthese studies. In a typical titration of A431 cell respiration it wasfound that 0.75 to 1.25 mg/ml of LYCD resulted in about a 60% increasein respiration. An LYCD concentration of 0.15 mg/ml gave a 10 to 15%increase in respiration; and 1.50 mg/ml of LYCD resulted in a 20%increase in respiration. Higher concentrations of LYCD resulted in nostimulation of baseline respiration.

Example 29

In companion experiments it was determined that 25 to 35 picomolarconcentrations of hydrocortisone inhibited A431 respiration, but, whenadded together with LYCD synergistically doubled the respiratorystimulation of 0.2 mg/ml LYCD from 20% to 40%.

A431 cell growth experiments were compared at seven days using astandard commercial serum-free medium (Gibco DME, Dulbecco's MinimumEssential media) and ITS (5 micrograms/ml each of insulin, transferinand selenium).

It was found that a 0.70 mg/ml of LYCD caused significant andreproducible growth of A431 cells. This concentration of LYCD representsabout 12% of the concentration of LYCD found in presently marketedproducts containing LYCD.

LYCD concentrations of 0.02 mg/ml provided a 30% enhancement in A431cell growth.

Hydrocortisone has been reported in the literature to be an inhibitor ofA431 cell growth.

Example 30

A seven day study was made to determine the effect of hydrocortisone oncell growth. It was found that hydrocortisone at the very lowconcentration of 1×10⁻⁸ mg./ml. inhibits A431 cell growth by 65%.However, it was further surprisingly found that the combination of thehydrocortisone plus 0.75 mg/ml of LYCD enhances cell growth by 200%.

Example 31

A LYCODERM®/hydrocortisone ointment combination product for topicalantiinflammatory therapy was formulated as follows:

    ______________________________________                                        Ingredient       Amount     % w/w                                             ______________________________________                                        Polysorbate 80   2 pounds   1.00                                              Hydrocortisone Acetate                                                                         1 pound    0.50                                              Phenyl Mercuric Nitrate                                                                        9.0 grams  0.01                                              LYCD (12 u/mg)   533.34 grams                                                                             1.0                                               Deionized water  2.25 pounds                                                                              2.0                                               Beeswax (white)  8 lb. 1 oz.                                                                              4.04                                              Lanolin          8 lb. 1 oz.                                                                              4.04                                              Petrolatum       168.5 pounds                                                                             84.28                                             Shark Liver Oil  6 lb. 1 oz.                                                                              3.03                                              Thyme Oil        90 grams   0.10                                              ______________________________________                                    

Procedure

The first 3 ingredients were combined and mixed well. The LYCD wasdissolved in water, combined with the first group and mixed well. Thethird group of ingredients was added to a clean container, heated to160° F. with good mixing. With stirring, the water mixture was heated to140° F. and added to the petrolatum preparation in a container. Whilestirring continued the mixture was cooled to 100° F. then the thyme oilwas added and the mixture further cooled to 60°-80° F. for filling. Theformulation thus prepared was found to provide a superior topicalantiinflammatory product.

Herpes zoster (shingles) is an acute inflammatory disease of thecerebral ganglia and ganglia of the posterior nerve roots, caused by thevirus of chicken pox. It is characterized by groups of small vesicles oninflammatory bases occurring in cutaneous areas supplied by certainnerve trunks, and associated with neuralgic pain.

Severe clinical herpes zoster is generally not helped by treatment withpresently available antiviral / anti-inflammatory medications.

Example 32

A number of herpes zoster patients were treated by Sidney Peerless, M.D.of E.N.T. Associates, 3131 Harvey avenue, Cincinnati Ohio 45229. Thetreatment was carried out after failure of conventional medication, andcomprised treatment with a composition according to the presentinvention comprising the LYCODERM®/Hydrocortisone Acetate formulationshown above in Example 31.

Patient 1: This patient had shingles of 4 weeks duration, herpes of theright face and forehead. Symptoms: severe pain, breaking out pustules,and redness. Previous treatment: Zovirex capsules, Zovirex ointment, andantibiotics did not help. The patient was placed onLYCODERM®/Hydrocortisone Acetate ointment of Example 31 applied 2-4times per day to the affected areas. In 3 days the patient showed markedimprovement, especially in the pustules and also in the pain threshold.Within 10 days the lesions were improved and the patient felt muchbetter symptomatically.

Patent 2: The patient had severe shingles in the cervical area goinginto the lower portion of the jaw and into the neck characterized bypustules, severe pain and erythema. Under previous treatment by adermatologist he had received steroids systemically and Zoviraxointment. After having the disease for three weeks he came to see Dr.Peerless in desperation, because of the severe pain he experienced. Thepatient also had herpes lesions in his throat. He was placed on theLYCD/Steroid combination ointment of Example 31. After 10 days oftreatment the entire facial and neck lesions were gone. There was amarked diminution of pain and need for Demerol, and his generalcondition improved greatly.

Patient 3: The patient had severe shingles involving the right posteriorleg and up to the dorsum of the foot. Under the care of anotherphysician the patient had received steroids, antibiotics systemically,and an antibiotic ointment, Polysporin, applied to the lesions withoutimprovement. The patient came to see Dr. Peerless also in desperation.She was placed on the LYCD/steroid combination ointment of Example 31.After three weeks the lesions were almost completely cleared. The painfactor was gone. The patient was able to wear her shoe and showedoverall marked improvement.

Patient 4: This patient had shingles of the lower lumbar area along thenerve root with large pustules, erythema and almost uncontrollable pain.He had been on pain medicine and Zovirax. He had also been givensystemic antibiotics and steroids with very little improvement. Thepatient was placed on the LYCODERM®/Hydrocortisone Acetate ointment ofExample 31. In 48 hours his condition improved markedly, especially inreduction of pain. After another 4-5 days on the medication the herpeticlesions were almost completely under control, and medication wascontinued for another week. Three weeks after stopping the medication,the patient had a recurrence of the herpes. Readministration of theLYCD/steroid ointment of Example 31 for two weeks again brought theherpes under control, and the patient has remained well.

Example 33

A formulation was analogously prepared in which the hydrocortisoneacetate concentration was reduced to 0.1%. This formulation was found toenhance the anti-erythema, wound-healing properties of the combinationproduct.

Example 34

Alternatively, LYCD at levels of 200 to 1800 units SRF per ounce(approximately 0.1% to 0.8%) are added to conventional formulations(creams, lotions, ointments, gels, etc.) of compatible topicaladrenocorticoid formulations which are used in concentrations of 0.01%to 1.0% to produce synergistic therapeutic compositions providing moreeffective medication for the same indications presently approved by theFDA.

A representative listing of Topical Adrenocorticoids which may be usedin formulating compositions according to the present invention maybefound in the U.S. Pharmacopeial Convention 1986 publication "THEPHYSICIANS' AND PHARMACISTS' GUIDE TO YOUR MEDICINES", published byBallantine Books, N.Y.N.Y.

Example 35

More particularly, the addition of 0.25% hydrocortisone acetate to theLYCODERN® formulation containing 1000 units of LYCD per ounce ofointment (approximately 0.50%) results in a topical antiinflammatorymedicinal composition with greater activity and efficacy than ispresently available in any topical steroid product licensed by the U.S.FDA for OTC (over the counter) use.

ANTI-SKIN WRINKLING COMPOSITIONS Example 36

Skin wrinkling is accelerated by deficiencies in collagensynthesis/metabolism. Vitamin A acid (all-trans retinoic acid) is usedas a topical preparation to slow or reverse the process of wrinkling.However, its use is limited by concentration related toxicity. Accordingto the present invention LYCD compositions synergistically augment theanti-wrinkling actions of topical retinoic acid with no increase intoxicity. Thus the LYCODERM® formulation previously described isprepared using 1000 units of SRF per ounce (ca 0.5% LYCD) and 0.25% ofvitamin A acid to provide a topical ointment composition synergisticallymore effective than retinoic acid in ameliorating the skin wrinklingprocess, including photo-aged skin.

Ecample 37

Alternatively, presently used cream (0.1%), gel (0.025%), or liquid(0.05%) Vitamin A acid formulations are augmented with 0.5% of LYCD(about 1000 units SRF per ounce to provide synergistically moreeffective anti-wrinkling compositions. Additionally, these novelcompositions are beneficial in treating ichthyosis, actinic keratosisand other hyperkeratotic conditions.

LYCD may also be combined with other retinoic acid congeners, knowncollectively as retinoids, which are also useful topical anti-wrinklingagents to produce new compositions as covered by the present invention.A number of these epidermally acting retinoids are described, forinstance, in the special issue supplement to The Journal of the AmericanAcademy of Dermatology (Volume 15, No. 4, Part 4, October 1986 entitled"TOPICAL RETINOIDS: AN UPDATE".

HAIR GROWTH STIMULATION COMPOSITIONS

Few effective agents are currently available for stimulation of hairgrowth. Only one topical preparation, a 2% solution of minoxidol,manufactured and marketed by the Upjohn Company, Kalamazoo, Mich., ispresently approved by the Food and Drug Administration as apharmaceutical preparation for the treatment of male pattern baldness(alopecia androgenica) of the vertex of the scalp. At least four monthsof continuous use is generally required before evidence of hair growthcan be seen. The historical development of topical treatments foralopecia is more fully set forth in U.S. Pat. Nos. 4,139, 619 and4,596,812 which are herein incorporated by reference.

It is an additional objective of the present invention to provide noveland effective treatments for male pattern baldness by the application oftopical pharmaceutical compositions incorporating LYCD at the 0.1% to3.0% level in pharmaceutically acceptable formulations (solutions,creams, gels, ointments, etc.).

Example 38

The following LYCD solution was prepared containing 2000 units of SRFper ounce:

    ______________________________________                                        LYCD/SRF            1.0%     w/v                                              Alcohol             55.0%    w/v                                              Oleyl alcohol       5.0%     w/v                                              Propylene glycol    to 100%                                                   ______________________________________                                    

When applied to the total affected are of the scalp of balding malestwice daily for four months, evidence of hair regrowth is observed.

Example 39

The following cream formulation of LYCD is also useful for promotinghair growth. The cream contains (per 100 grams):

    ______________________________________                                        LYCD/SRF               5250    units                                          Dimethicone            5.0     g                                              D-pantheol             4.0     g                                              Benzalkonium Chloride  0.1     g                                              A water washable cream base                                                                          to 100  g                                              ______________________________________                                    

Example 40

For some individuals, another effective composition for treating malepattern alopecia combines LYCODERM® ointment and very low dosehydrocortisone acetate, formulated as follows:

    ______________________________________                                        Ingredient         per 100 parts by wt.                                       ______________________________________                                        Polysorbate 80     1.0                                                        Hydrocortisone Acetate                                                                           0.1                                                        Phenyl Mercuric Nitrate                                                                           0.01                                                      Live Yeast Cell Derivative                                                                       1.0                                                        (LYCD/SRF, 2000 units/oz.)                                                    (approximate)                                                                 Deionized water    2.0                                                        Beeswax (white)    4.0                                                        Lanolin            4.0                                                        Petrolatum         84.8                                                       Shark Liver Oil    3.0                                                        Thyme Oil          0.1                                                        ______________________________________                                    

Procedure

The first three ingredients were combined and mixed well. The LYCD wasdissolved in water, added to the first group and again mixed well. Thethird group of ingredients were added separately to a clean containerand heated to 160° F. with good mixing. With continued stirring, thewater solution was heated to 140° F. and added to thepetrolatum/lanolin/beeswax mixture. Stirring continued as the totalpreparation was allowed to cool to 100° F. Then the thyme oil wasstirred in and the mixture further cooled to 60°-80° F. for filling.

Example 41

Combinations of LYCD with low doses of Minoxidol or retinoic acid alsoprovide synergistic compositions for treating alopecia androgenica.Advantageously, 0.5% of LYCD is formulated with 1.0% minoxidol solutionor with 0.1% of vitamin A acid to provide more effective hair growthcompositions, although application for four months may still be requiredbefore evidence of hair regrowth is observed.

COMPOSITIONS FOR USE IN OPHTHALMOLOGY Example 42

A 1% to 10% solution of LYCD, in a pharmaceutically acceptableophthalmic formulation provides a useful composition for the treatmentof various eye problems and accelerates corneal epithelial regenerationand the healing of stromal incisions following corneal transplantsurgery. More particularly, a 5% solution of LYCD is useful for theacceleration of corneal epithelial regeneration and the healing ofstromal tissue in the condition of non-healing corneal defects.

Formulated in combination with low doses of a fibronectin or a laminin(one to 100 nanograms per ml.) LYCD is synergistically more effective intreating Dry Eye, Corneal Incisions, Recurrent Corneal Erosions, andNon-healing Corneal Defects.

Example 43

For the treatment of ocular viral infections, such as, for instance,keratitis due to Herpes simplex virus, the 5% solution of LYCD, in apharmaceutically acceptable ophthalmic formulation is augmented with anantiviral agent (i.e. 1% trifuridine or acyclovir) and 0.5%hydrocortisone (as its water soluble ester).

The compositions of the present invention comprising LYCD in combinationwith other topically active medicinal ingredients have many advantagesover conventional products. The presence of the LYCD in the compositionsprovides a synergistic effect which makes the conventional materialsmore effective and permits less of the conventional ingredients to beused while still achieving the same results.

The compositions of the present invention contain, in addition to LYCDand the other pharmaceutically active ingredient, a carrier suitable forrendering the composition as a formulation to be used for topicalapplications. In one method for forming the compositions of the presentinvention the carrier is provided by the LYCODERM®. In another methodthe carrier is provided by the commercial form of the other activeingredient. Alternatively, a suitable carrier known in the art may beadded to both the LYCD and the other active ingredient. In all examplesabove the indicated percent content of the stated ingredients is basedon the weight of the total ingredients, the LYCD, the otherpharmaceutically active ingredient, and the carrier.

The herein described new topical compositions and methods of treatmentfor a variety of skin conditions are equally applicable to veterinaryproblems, that is, the treatment of farm animals as well as domesticpets.

Although the invention has been described in connection with specificembodiments thereof, it is evident that many alternatives,modifications, and variations will be apparent to those skilled in theart in the light of the foregoing description. Accordingly, it isintended to embrace all such alternatives, modifications and variationswithin the spirit and scope of the invention as defined by the appendedclaims.

Invention is claimed as follows:
 1. A topical composition adapted forapplication to the skin comprising in admixture with a pharmaceuticallyacceptable topical carrier, an antiinfective agent effective in thetreatment of acne, and from about 0.1% to about 3.0% by weight of saidcomposition of Live Yeast Cell Derivative (LYCD) in amounts effective toameliorate the effects of acne when applied to an affected area of theskin.
 2. A topical composition according to claim 1, wherein said LYCDis present in an amount of from about 0.1% to about 1.0% by weight ofsaid composition.
 3. A topical composition according to claim 2, whereinsaid antiinfective agent is erythromycin, or a pharmaceutically usefulester thereof.
 4. A topical composition according to claim 2, whereinsaid antiinfective agent is lincomycin hydrochloride.
 5. A topicalcomposition according to claim 2, wherein said antiinfective agent isclindamycin phosphate.
 6. A topical composition according to claim 2,wherein said antiinfective agent is benzoyl peroxide.
 7. A topicalcomposition according to claim 2, wherein said antiinfective agent isvitamin A acid.
 8. A method for ameliorating the topical symptomsassociated with acne infection in a human being, which comprisestopically applying to an area of the skin of said human being where thesymptoms are manifested an effective amount of a composition of claim 1.9. A method according to claim 8, wherein antiinfective agent iserythromycin or a pharmaceutically useful ester thereof.
 10. A methodaccording to claim 8, wherein said antiinfective agent is lincomycinhydrochloride.
 11. A method according to claim 8, wherein saidantiinfective agent is clindamycin phosphate.
 12. A method according toclaim 8, wherein said antiinfective agent is benzoyl peroxide.
 13. Amethod according to claim 8, wherein said antiinfective agent is vitaminA acid.